rabbit anti stat2 total  (Novus Biologicals)

Journal: Viruses

Article Title: Human Platelet Lysate Induces Antiviral Responses against Parechovirus A3

doi: 10.3390/v14071499

Figure Lengend Snippet: hPL-induced innate immunity response type I interferon and antiviral-related genes on GBM cells. ( A ) GBM cells were cultured under FBS, 2.5% and 5% hPL for 6 h, 24 h, and 48 h. The cell protein lysates were employed to analyze the innate immunity pathways by immunoblotting using anti-phospho-STAT1, anti-total STAT2, anti-phospho-STAT2, anti-total STAT2, anti-phospho-IRF3, anti-total IRF3, anti-phospho-NFκB, anti-total NFκB p65, and anti-GAPDH. ( B ) Quantification of protein expressions was normalized to GAPDH. Data are mean ± SD of three independent experiments. Student’s t -test, *, p < 0.05; **, p <0.01 compared with GBM cells supplemented with FBS. ( C ) GBM cells were cultured under FBS, 2.5% and 5% hPL mediums for 24 h and 48 h. The mRNA level of antiviral-related genes IFN-α , IFN-β , IRF3 , and MxA were confirmed by qPCR. The level of transcripts was normalized to GAPDH. Data are mean ± SD of three independent experimental samples. Student’s t -test, *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with FBS.

Article Snippet: The antibodies were anti-PeV VP0 (LTK BioLaboratories, Taoyuan, Taiwan) [ , ], anti-phospho STAT2 (Cell Signaling, 88410, Danvers, MA, USA), anti-total STAT2 (Cell Signaling, #72604), anti-phospho STAT1 (Cell Signaling, #9167), anti-total STAT1 (Cell Signaling, #14994), anti-phospho IRF3 (Abcam, #ab76493, Cambridge, England), anti-total IRF3 (Cell Signaling, #11904), anti-phospho NFκB p65 (Abcam, #ab86299), anti-total NFκB p65 (Cell Signaling, #8242) and anti-GAPDH (Proteintech, 60004-1-Ig, Rosemont, IL, USA).

Techniques: Cell Culture, Western Blot

Journal: Viruses

Article Title: Human Platelet Lysate Induces Antiviral Responses against Parechovirus A3

doi: 10.3390/v14071499

Figure Lengend Snippet: hPL-supplemented growth medium-induced expression of IFN-signaling pathway under PeV-A3 infection of GBM cells. ( A ) GBM cells were infected with PeV-A3 at MOI = 1 for 6 h, 24 h, and 48 h under FBS- or hPL-supplemented growth mediums. Cellular protein lysates were extracted to analyze IFN-signaling pathway by immunoblotting using anti-PeV VP0, anti-phospho-STAT1, anti-total STAT1, anti-phospho-STAT2, anti-total STAT2, anti-phospho-IRF3, anti-total IRF3, anti-phospho-NFκB, anti-total NFκB p65, and anti-GAPDH. ( B ) Quantification of different protein expressions was normalized to GAPDH. Data are mean ± SD of three independent samples. Student’s t -test, *, p < 0.05 compared with FBS.

Article Snippet: The antibodies were anti-PeV VP0 (LTK BioLaboratories, Taoyuan, Taiwan) [ , ], anti-phospho STAT2 (Cell Signaling, 88410, Danvers, MA, USA), anti-total STAT2 (Cell Signaling, #72604), anti-phospho STAT1 (Cell Signaling, #9167), anti-total STAT1 (Cell Signaling, #14994), anti-phospho IRF3 (Abcam, #ab76493, Cambridge, England), anti-total IRF3 (Cell Signaling, #11904), anti-phospho NFκB p65 (Abcam, #ab86299), anti-total NFκB p65 (Cell Signaling, #8242) and anti-GAPDH (Proteintech, 60004-1-Ig, Rosemont, IL, USA).

Techniques: Expressing, Infection, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Contribution of the STAT Family of Transcription Factors to the Expression of the Serotonin 2B (HTR2B) Receptor in Human Uveal Melanoma

doi: 10.3390/ijms23031564

Figure Lengend Snippet: Western blot analysis of the STAT isoforms in UM cells. Total proteins isolated from the UM cell lines T97, T108, T142 and T143 were Western blotted using polyclonal antibodies directed against all STAT isoforms (STAT1 to STAT6). Actin was also blotted as a control. Coomassie blue staining (25 μg of each protein extract was used) is also shown beside the Western blots as a protein loading control. UM cell lines were found to express all STAT proteins to varying levels. In addition, the metastatic T142 cell line expresses a STAT2 isoform with an apparent molecular mass higher than that observed in the remaining UM cells.

Article Snippet: Western blots were conducted as described [ , , , , ] using antibodies directed against the following proteins: Total STAT1 (D1K9Y (polyclonal), 1/300; Cell Signaling Technology), Total STAT2 (D9J7L (polyclonal), 1/300; Cell Signaling Technology, Danvers, MA, USA), Total STAT3 (sc-8019 (monoclonal), 1/100; Santa Cruz Biotechnology, Dallas, TX, USA), Total STAT4 (sc-398228 (monoclonal), 1/100; Santa Cruz Biotechnology), Total STAT5 (sc-74442 (monoclonal), 1/100; Santa Cruz Biotechnology), Total STAT6 (sc-374021 (monoclonal), 1/100; Santa Cruz Biotechnology), HTR2B (HPA-012867 (monoclonal), 1/100; Millipore Sigma), Phospho STAT1 (sc-8394 (monoclonal), 1/100; Santa Cruz Biotechnology), Phospho STAT3 (sc-8059 (monoclonal), 1/100; Santa Cruz Biotechnology), Phospho STAT5 (sc-81524 (monoclonal), 1/100; Santa Cruz Biotechnology), and a peroxidase-conjugated AffiniPure Goat secondary antibody against mouse IgG (1:2500 dilution; Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Western Blot, Isolation, Staining